RESUMO
B-cells have received little attention in axial spondyloarthritis (axSpA) and for this reason their role in pathogenesis remains unclear. However, there are indications that B-cells may be involved in the disease process. Our objective was to obtain insights into the composition of the peripheral B-cell compartment of axSpA patients compared to healthy donors (HD) and patients with primary Sjögren's syndrome (pSS), a typical B-cell-associated autoimmune disease. Special emphasis was given to CD27-negative B-cells expressing low levels of CD21 (CD21low B-cells), since this subset is implicated in autoimmune diseases with strong involvement of B-cells. Transitional B-cells (CD38hi) were excluded from the analysis of the CD27-CD21low B-cell compartment. This study included 45 axSpA patients, 20 pSS patients and 30 HDs. Intriguingly, compared to HDs the frequency of CD27-CD38lowCD21low B-cells was significantly elevated in both axSpA and pSS patients (P<0.0001 for both comparisons). The frequency of CD27-CD38lowCD21low B-cells expressing the activation-induced immune markers T-bet and CD11c was decreased in axSpA patients compared to HDs. A higher proportion of CD27-CD38lowCD21low B-cells expressed the chemokine receptor CXCR3 in axSpA compared to HDs, suggestive for active involvement of these cells in an inflammatory process. The frequency of CD27-CD38lowCD21low B-cells in axSpA patients correlated positively with age and erythrocyte sedimentation rate. Furthermore, axSpA patients with extra-skeletal manifestations (ESM) showed increased frequencies of CD27-CD38lowCD21low B-cells compared to patients without ESM. In conclusion, our findings are suggestive of active B-cell involvement in the pathogenesis of axSpA, against prevailing dogma.
Assuntos
ADP-Ribosil Ciclase 1/sangue , Linfócitos B/imunologia , Síndrome de Sjogren/imunologia , Espondilartrite/imunologia , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/sangue , Adulto , Linfócitos B/metabolismo , Biomarcadores/sangue , Antígeno CD11c/sangue , Estudos de Casos e Controles , Feminino , Citometria de Fluxo , Humanos , Masculino , Pessoa de Meia-Idade , Receptores de Complemento 3d/sangue , Síndrome de Sjogren/sangue , Síndrome de Sjogren/diagnóstico , Espondilartrite/sangue , Espondilartrite/diagnósticoRESUMO
Circulating CD11c+ B cells are a key phenomenon in certain types of autoimmunity but have also been described in the context of regular immune responses (i.e., infections, vaccination). Using mass cytometry to profile 46 different markers on individual immune cells, we systematically initially confirmed the presence of increased CD11c+ B cells in the blood of systemic lupus erythematosus (SLE) patients. Notably, significant differences in the expression of CD21, CD27, and CD38 became apparent between CD11c- and CD11c+ B cells. We observed direct correlation of the frequency of CD21-CD27- B cells and CD21-CD38- B cells with CD11c+ B cells, which were most pronounced in SLE compared to primary Sjögren's syndrome patients (pSS) and healthy donors (HD). Thus, CD11c+ B cells resided mainly within memory subsets and were enriched in CD27-IgD-, CD21-CD27-, and CD21-CD38- B cell phenotypes. CD11c+ B cells from all donor groups (SLE, pSS, and HD) showed enhanced CD69, Ki-67, CD45RO, CD45RA, and CD19 expression, whereas the membrane expression of CXCR5 and CD21 were diminished. Notably, SLE CD11c+ B cells showed enhanced expression of the checkpoint molecules CD86, PD1, PDL1, CD137, VISTA, and CTLA-4 compared to HD. The substantial increase of CD11c+ B cells with a CD21- phenotype co-expressing distinct activation and checkpoint markers, points to a quantitative increased alternate (extrafollicular) B cell activation route possibly related to abnormal immune regulation as seen under the striking inflammatory conditions of SLE which shows a characteristic PD-1/PD-L1 upregulation.
Assuntos
Autoimunidade , Linfócitos B/imunologia , Antígeno CD11c/sangue , Citometria de Fluxo , Imunofenotipagem , Lúpus Eritematoso Sistêmico/imunologia , Ativação Linfocitária , Síndrome de Sjogren/imunologia , ADP-Ribosil Ciclase 1/sangue , Linfócitos B/metabolismo , Antígeno B7-H1/sangue , Biomarcadores/sangue , Estudos de Casos e Controles , Humanos , Lúpus Eritematoso Sistêmico/sangue , Lúpus Eritematoso Sistêmico/diagnóstico , Glicoproteínas de Membrana/sangue , Fenótipo , Receptor de Morte Celular Programada 1/sangue , Receptores de Complemento 3d/sangue , Síndrome de Sjogren/sangue , Síndrome de Sjogren/diagnóstico , Membro 7 da Superfamília de Receptores de Fatores de Necrose Tumoral/sangueRESUMO
BACKGROUND: Epstein-Barr virus (EBV) DNAemia is a major risk factor for posttransplant lymphoproliferative disorder; however, immune correlates of EBV DNAemia in the transplant setting are limited. METHODS: Peripheral blood mononuclear cells were collected from 30 transplant recipients with self-limiting EBV DNAemia (SLD; n = 11) or chronic EBV DNAemia (CD; n = 19) at enrollment and 4-8 weeks later. Mass cytometry was used to characterize innate and T-cell immune correlates of EBV DNAemia. Furthermore, flow cytometry was used to measure the frequency of EBV-specific T-cell responses between groups following stimulation with an EBV-infected cell lysate. RESULTS: Unsupervised analysis of the innate compartment (CD3CD19 cells) identified 5 CD11c clusters at higher abundance in the SLD group (false discovery rate ≤ 1%). These clusters expressed CD11b, CD45RO, CD14, CD123, CD127, and CD38, among others. Unsupervised profiling of the T-cell compartment (CD3CD19) revealed 2 CD4 T-cell clusters at higher frequency among those with SLD (false discovery rate ≤ 1%), which expressed CD45RA, CCR7, CD27, CD28, and CD40L-suggestive of a naive T cell (TN). Manual biaxial gating confirmed increased frequencies of conventional dendritic cells (3.1% versus 2.1%; P = 0.023) and CD4 TN (4.4% versus 1.9%; P = 0.018) among those with SLD. Last, frequencies of interferon-γ-producing EBV-specific CD4 T cells were significantly lower in the CD group relative to those with SLD (4243 versus 250 cells/10 cells; P = 0.015). CONCLUSIONS: CD is associated with a reduction of CD11c cells, CD4 TN, and interferon-γ-producing EBV-specific CD4 T cells, suggesting an interplay between innate and adaptive immune compartments may be important for regulating EBV DNAemia.
Assuntos
Imunidade Adaptativa , Linfócitos T CD4-Positivos/imunologia , DNA Viral/sangue , Células Dendríticas/imunologia , Infecções por Vírus Epstein-Barr/imunologia , Herpesvirus Humano 4/imunologia , Imunidade Inata , Infecções Oportunistas/imunologia , Transplante de Órgãos/efeitos adversos , Adulto , Idoso , Antígeno CD11c/sangue , Linfócitos T CD4-Positivos/metabolismo , Linfócitos T CD4-Positivos/virologia , Doença Crônica , Estudos Transversais , Células Dendríticas/metabolismo , Células Dendríticas/virologia , Infecções por Vírus Epstein-Barr/sangue , Infecções por Vírus Epstein-Barr/diagnóstico , Infecções por Vírus Epstein-Barr/virologia , Feminino , Herpesvirus Humano 4/genética , Interações Hospedeiro-Patógeno , Humanos , Interferon gama/sangue , Masculino , Pessoa de Meia-Idade , Infecções Oportunistas/sangue , Infecções Oportunistas/diagnóstico , Infecções Oportunistas/virologia , Resultado do Tratamento , Carga Viral , Adulto JovemRESUMO
Endothelial dysfunction and inflammation are key mechanisms of vascular disease. We hypothesised that heterogeneity of monocyte subpopulations may be related to the development of vascular dysfunction in coronary artery disease (CAD). Therefore, we examined the relationships between monocyte subsets (CD14++CD16- "classical - Mon1", CD14++CD16+ "intermediate - Mon2" and CD14+CD16++ "nonclassical - Mon3"), endothelial function and risk factor profiles in 130 patients with CAD undergoing coronary artery bypass grafting. This allowed for direct nitric oxide (NO) bioavailability assessment using isometric tension studies ex vivo (acetylcholine; ACh- and sodium-nitropruside; SNP-dependent) in segments of internal mammary arteries. The expression of CD14 and CD16 antigens and activation markers were determined in peripheral blood mononuclear cells using flow cytometry. Patients with high CD14+CD16++ "nonclassical" and low CD14++CD16- "classical" monocytes presented impaired endothelial function. High frequency of CD14+CD16++ "nonclassical" monocytes was associated with increased vascular superoxide production. Furthermore, endothelial dysfunction was associated with higher expression of activation marker CD11c selectively on CD14+CD16++ monocytes. Nonclassical and classical monocyte frequencies remained independent predictors of endothelial dysfunction when major risk factors for atherosclerosis were taken into account (ß=0.18 p=0.04 and ß=-0.19 p=0.03, respectively). In summary, our data indicate that CD14+CD16++ "nonclassical" monocytes are associated with more advanced vascular dysfunction measured as NO- bioavailability and vascular reactive oxygen species production.
Assuntos
Doença da Artéria Coronariana/sangue , Doença da Artéria Coronariana/fisiopatologia , Endotélio Vascular/fisiopatologia , Receptores de Lipopolissacarídeos/sangue , Artéria Torácica Interna/fisiopatologia , Monócitos/metabolismo , Receptores de IgG/sangue , Vasodilatação , Idoso , Biomarcadores/sangue , Antígeno CD11c/sangue , Ponte de Artéria Coronária , Doença da Artéria Coronariana/diagnóstico , Doença da Artéria Coronariana/imunologia , Endotélio Vascular/metabolismo , Feminino , Proteínas Ligadas por GPI/sangue , Humanos , Masculino , Artéria Torácica Interna/metabolismo , Pessoa de Meia-Idade , Monócitos/imunologia , Óxido Nítrico/metabolismo , Fenótipo , Superóxidos/metabolismoRESUMO
INTRODUCTION: Gene expression profiling is rapidly becoming a useful and informative tool in a much needed area of research. Identifying patients as to whether they will respond or not to a given treatment before prescription is not only essential to optimise treatment outcome but also to lessen the economic burden that such drugs can have on healthcare resources. In rheumatoid arthritis (RA), there is of yet no genetic/genomic biomarker which can accurately predict response to TNF inhibitor biologics prior to treatment, despite much interest in this area. Multiple studies have reported findings on potential candidate genes; however, due to relatively small sample sizes or lack of sufficient validation, results have been disappointingly inconsistent. The aim of this research was to further explore the predictive value of a previously reported association between CD11c expression and response to the TNF inhibitor biologics, adalimumab and etanercept. METHODS: Real-time qPCR was performed using whole blood RNA samples obtained from seventy-five rheumatoid arthritis patients about to commence treatment with a TNF inhibitor biologic drug, whose response status was determined at 3-month follow-up using the EULAR classification criteria. Relative quantification of CD11c using the comparative CT method outputted differential expression between good-responders and non-responders as a fold-change. RESULTS: Relative expression of CD11c in patients receiving TNF inhibitor biologics yielded a decrease of 1.025 fold in good-responders as compared to non-responders (p-value = 0.36). Upon stratification of patients dependent upon the specific drug administered, adalimumab or etanercept, similar findings to the full cohort were observed, decreases of 1.015 (p-value = 0.33) and 1.032 fold (p-value = 0.13) in good-responders compared to non-responders, respectively. CONCLUSION: The results from this study reveal that CD11c expression does not correlate with response to TNF inhibitor biologics when tested for within pre-treatment whole blood samples of rheumatoid arthritis patients.
Assuntos
Antirreumáticos/uso terapêutico , Artrite Reumatoide/sangue , Artrite Reumatoide/tratamento farmacológico , Biomarcadores/sangue , Antígeno CD11c/sangue , Adalimumab/uso terapêutico , Produtos Biológicos/uso terapêutico , Etanercepte/uso terapêutico , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase em Tempo Real , Resultado do Tratamento , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
Autoimmune diseases develop when self-specific T cells that escaped negative selection initiate a harmful immune response against self. However, factors, which influence the initiation and progression of an autoimmune response remain incompletely understood. By establishing a double-transgenic BALB/c mouse system in which different amounts of a cell-surface neo-self-antigen are expressed under the CD11c promoter, we demonstrate that antigen dose dramatically influences T-cell tolerance mechanisms. Moderate antigen expression in both hematopoietic and nonhematopoietic cells favors the development of antigen-specific Treg cells and the establishment of a tolerogenic environment. In marked contrast, a high dose of antigen expression results in very stringent negative selection, in poor development of antigen-specific Treg cells and in the early onset of anemia and splenomegaly and the late development of arthritis and high titers of IgG auto Abs. Disease is initiated by autoreactive T cells, which escape negative selection by expressing a second TCR with a different specificity or an altered affinity. Transfer of Ag-specific Treg cells ameliorates the early onset signs of disease but does not prevent the development of long-term chronic pathologies. Altogether, our results suggest that Ag dose directly affects Treg-cell generation and thus, the set-up of T-cell tolerance.
Assuntos
Autoantígenos/imunologia , Doenças Autoimunes/imunologia , Imunoglobulina G/imunologia , Linfócitos T Reguladores/imunologia , Animais , Autoanticorpos/sangue , Autoanticorpos/imunologia , Autoantígenos/sangue , Autoantígenos/genética , Doenças Autoimunes/sangue , Doenças Autoimunes/genética , Doenças Autoimunes/patologia , Antígeno CD11c/sangue , Antígeno CD11c/genética , Antígeno CD11c/imunologia , Relação Dose-Resposta Imunológica , Tolerância Imunológica , Imunoglobulina G/sangue , Imunoglobulina G/genética , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Transgênicos , Receptores de Antígenos de Linfócitos T , Linfócitos T Reguladores/metabolismo , Linfócitos T Reguladores/patologiaRESUMO
In an anti-glomerular basement membrane (GBM) glomerulonephritis (GN) model, GN-resistant Lewis rats naturally recover from early glomerular inflammation. Here we investigated recovery mechanisms for development of a potential immunotherapy for autoimmune GN. Our previous studies suggested that glomeruli-infiltrating leukocytes with a phenotype of CD8αα+CD11c+MHC-II+CD3- (GIL CD8αα+ cells) were responsible for recovery through induction of T-cell apoptosis. Now, we identified peripheral blood CD8αα+CD11c+MHC-II+CD3- cells (PBMC CD8αα+CD3- cells), which shared 9 markers with GIL CD8αα+ cells. Upon incubation, PBMC CD8αα+CD3- cells displayed a morphology resembling that of dendritic cells. Similar to GIL CD8αα+ cells, PBMC CD8αα+CD3- cells were capable of inducing T-cell apoptosis in vitro. Hence, PBMC CD8αα+CD3- cells were likely the precursor of GIL CD8αα+ cells. We next tested their potential in vivo function. PBMC CD8αα+CD3- cells were able to infiltrate inflamed but not normal glomeruli. Isolated PBMC CD8αα+CD3- cells of Lewis rats were transferred into GN-prone Wistar-Kyoto rats at early inflammatory stage (days 17-25). When examined at day 45, both histopathology and blood urea nitrogen/serum creatinine level showed significantly attenuated GN in 80% of cell recipient Wistar-Kyoto rats. Separate experiments verified infiltration of transferred Lewis PBMC CD8αα+CD3- into the glomeruli, accompanied with apoptotic CD4+ T cells in the glomeruli of the recipient Wistar-Kyoto rats. Thus, PBMC CD8αα+CD3- cells of Lewis rats were able to terminate ongoing autoimmune inflammation in the glomeruli.
Assuntos
Transferência Adotiva , Autoimunidade , Antígeno CD11c/sangue , Complexo CD3/sangue , Antígenos CD8/sangue , Linfócitos T CD8-Positivos/transplante , Glomerulonefrite/prevenção & controle , Antígenos de Histocompatibilidade Classe II/sangue , Glomérulos Renais/imunologia , Animais , Apoptose , Biomarcadores/sangue , Nitrogênio da Ureia Sanguínea , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/patologia , Linfócitos T CD8-Positivos/imunologia , Células Cultivadas , Quimiotaxia de Leucócito , Técnicas de Cocultura , Creatinina/sangue , Citotoxicidade Imunológica , Modelos Animais de Doenças , Feminino , Adjuvante de Freund , Glomerulonefrite/sangue , Glomerulonefrite/induzido quimicamente , Glomerulonefrite/imunologia , Glomerulonefrite/patologia , Glomérulos Renais/patologia , Técnicas de Cultura de Órgãos , Peptídeos , Ratos Endogâmicos Lew , Ratos Endogâmicos WKY , Fatores de TempoRESUMO
OBJECTIVES: To examine the utility of CD11c expression on monocytes in normal controls and patients with chronic myelomonocytic leukemia (CMML) (n = 23) with flow cytometric immunophenotyping. METHODS: Twenty-three CMML samples and 10 control bone marrows submitted for lymphoma staging without evidence of disease were examined. RESULTS: Monocytes in CMML samples ranged from 4% to 35%. Expression of at least one aberrant monocytic marker was found on the monocytes in 18 (82%) of 22 evaluable cases. The most common aberrancy was underexpression of CD11c (n = 15), while none of the bone marrow controls showed underexpression of CD11c. CONCLUSIONS: A distinct heterogeneous population of monocytic cells with underexpression of CD11c was identified in all these cases. CD11c underexpression was independent of other aberrancies, including HLA-DR underexpression (n = 14), aberrant CD56 expression (n = 11), and underexpression of CD33, CD38, and CD14 (n = 6, 5, and 5, respectively), supporting the utility of CD11c expression status on monocytes in establishing a CMML diagnosis.
Assuntos
Antígeno CD11c/sangue , Leucemia Mielomonocítica Crônica/patologia , Monócitos/patologia , Antígenos CD/sangue , Biomarcadores/metabolismo , Células da Medula Óssea/metabolismo , Células da Medula Óssea/patologia , Feminino , Antígenos HLA-DR/sangue , Humanos , Leucemia Mielogênica Crônica BCR-ABL Positiva/complicações , Leucemia Mielogênica Crônica BCR-ABL Positiva/metabolismo , Leucemia Mielogênica Crônica BCR-ABL Positiva/patologia , Leucemia Mielomonocítica Crônica/metabolismo , Leucocitose/complicações , Leucocitose/metabolismo , Leucocitose/patologia , Masculino , Monócitos/metabolismo , Estadiamento de NeoplasiasRESUMO
BACKGROUND AND AIM: In human blood, two main subsets of antigen-presenting-cells (APCs) have been described: plasmocytoid dendritic cells (pDC) and myeloid dendritic cells (mDC) which are further subdivided in CD11c-mDC and CD16-mDC DC. In ulcerative colitis patients (UC) peripheral blood APCs express significant levels of the activation and lack immature-tolerogeneic APCs. Adacolumn selective granulocytapheresis (GCAP) has been associated with clinical efficacy in patients with UC. In the present study we sought the effect of sequential GCAP procedures in peripheral blood APCs in patients with UC and the effect on soluble cytokines. METHODS: We used multiparametric flow cytometry to quantify peripheral blood APCs and serum cytokines in 210 samples obtained from seven patients with steroid-dependent or steroid resistant UC undergoing GCAP treatment. Samples were drawn before, after 30 and 60 min of each session. RESULTS: Each GCAP session resulted in a dramatic tenfold reduction of peripheral blood CD16-mDC (P < 0.01), pDC decreased twofold (P = 0.05) but CD11c-mDC remained unchanged. This depletion was reached after 30 min and maintained at 60 min. The depletion of CD16-mDC and monocytes was associated with a reduction of serum tumor necrosis factor levels and a raise in interleukin-10 levels, although no statistical difference was reached. CONCLUSION: The effect of GCAP in peripheral blood APC consisted mainly on a significant depletion of tumor necrosis factor-α secreting CD16-mDC. This finding could suggest a potential mechanism of GCAP beneficial effect that must be confirmed in larger series.
Assuntos
Colite Ulcerativa/terapia , Células Dendríticas/imunologia , Granulócitos/imunologia , Mediadores da Inflamação/metabolismo , Leucaférese , Receptores de IgG/sangue , Fator de Necrose Tumoral alfa/sangue , Adolescente , Adulto , Idoso , Antígeno CD11c/sangue , Estudos de Casos e Controles , Colite Ulcerativa/imunologia , Feminino , Citometria de Fluxo , Proteínas Ligadas por GPI/sangue , Humanos , Contagem de Leucócitos , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Espanha , Resultado do Tratamento , Adulto JovemRESUMO
We performed transcription profiling using monocytes to identify predictive markers of response to anti-tumor necrosis factor (anti-TNF) therapy in patients with rheumatoid arthritis (RA). Several potential predictors of response were identified, including CD11c. Validation in samples from independent cohorts (total of n = 27 patients) using reverse transcription-PCR confirmed increased expression of CD11c in responders to adalimumab (100% sensitivity; 91.7% specificity, power 99.6%; alpha = 0.01). Pretherapy CD11c levels significantly correlated with the response criteria as defined by the American College of Rheumatology (ACR) (r = 0.656, P < 0.0001). However, CD11c was neither predictive of response to methotrexate (MTX) alone (n = 34) nor to MTX in combination with adalimumab (n = 16). Clinical responders revealed a reset to a normal expression pattern of resident/inflammatory monocyte markers, which was absent in nonresponders. Therefore, an analysis of key cell types identifies potentially predictive biomarkers that may help to restrict the use of adalimumab to therapy responders. Larger studies, including studies of monotherapy with other drugs, are now needed to confirm and validate the specificity of CD11c for anti-TNF biologics.
Assuntos
Anticorpos Monoclonais/sangue , Antirreumáticos/sangue , Artrite Reumatoide/sangue , Antígeno CD11c/sangue , Antígeno CD11c/genética , Transcrição Gênica/fisiologia , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Adalimumab , Adulto , Idoso , Anticorpos Monoclonais/fisiologia , Anticorpos Monoclonais/uso terapêutico , Anticorpos Monoclonais Humanizados , Antirreumáticos/farmacologia , Antirreumáticos/uso terapêutico , Artrite Reumatoide/tratamento farmacológico , Artrite Reumatoide/genética , Biomarcadores/sangue , Estudos de Coortes , Feminino , Marcadores Genéticos/genética , Humanos , Masculino , Metotrexato/uso terapêutico , Pessoa de Meia-Idade , Monócitos/efeitos dos fármacos , Monócitos/metabolismo , Valor Preditivo dos Testes , Análise Serial de Proteínas/métodos , Transcrição Gênica/efeitos dos fármacos , Resultado do Tratamento , Fator de Necrose Tumoral alfa/imunologiaRESUMO
Dendritic cells (DC) are antigen-presenting cells involved in induction and regulation of immune responses. We investigated the impact of the number of infused and day 28 dendritic cells on the development of acute and chronic GVHD (aGVHD, cGVHD). Monocytoid (MC) and plasmacytoid (PC) dendritic cells were characterized as lin(-)HLA-DR(+)CD11c(+) and lin(-)HLA-DR(+)CD123(+), respectively. Sixty-eight consecutive patients who underwent HLA matched related granuloyte-colony stimulating factor (G-CSF) mobilized allogeneic PBSCT, from February 2005 to May 2006, were included in the analysis. Twenty-three patients developed aGVHD (grade II-IV) and 21 patients had cGVHD. On a univariate analysis the day 28 total DC and the day 28 MC and PC dendritic cells as continuous variables were significantly associated with development of aGVHD and cGVHD. Using an ROC plot analysis a cutoff value for total DC = 10.7/microL, MC = 9.7/microL, and PC = 4.5/microL on day 28 gave the highest likelihood ratios for aGVHD (2.7, 2.14, and 3.29, respectively). On a multivariate analysis, a low day 28 PC (Assuntos
Antígeno CD11c/sangue
, Células Dendríticas
, Doença Enxerto-Hospedeiro/sangue
, Antígenos HLA-DR/sangue
, Subunidade alfa de Receptor de Interleucina-3/sangue
, Contagem de Leucócitos
, Transplante de Células-Tronco de Sangue Periférico
, Plasmócitos
, Receptores de Interleucina-3/sangue
, Doença Aguda
, Adolescente
, Adulto
, Criança
, Pré-Escolar
, Doença Crônica
, Células Dendríticas/patologia
, Feminino
, Doença Enxerto-Hospedeiro/etiologia
, Doença Enxerto-Hospedeiro/mortalidade
, Doença Enxerto-Hospedeiro/patologia
, Fator Estimulador de Colônias de Granulócitos/administração & dosagem
, Doenças Hematológicas/sangue
, Doenças Hematológicas/complicações
, Doenças Hematológicas/mortalidade
, Doenças Hematológicas/terapia
, Mobilização de Células-Tronco Hematopoéticas
, Teste de Histocompatibilidade
, Humanos
, Incidência
, Masculino
, Pessoa de Meia-Idade
, Transplante de Células-Tronco de Sangue Periférico/mortalidade
, Plasmócitos/patologia
, Valor Preditivo dos Testes
, Fatores de Risco
, Fatores de Tempo
, Transplante Homólogo
RESUMO
Dendritic cell (DC) immunotherapy for cancer has shown promising results in phase I and II clinical trials. Most studies have used monocyte-derived DCs (MoDCs) but their poor migratory capacity in vivo has emerged as a key issue. The natural circulating peripheral blood CD11c+ DC precursors (BDCs) may be an attractive alternative to MoDCs, as they can be isolated rapidly in sufficient quantities, and have superior migratory and T helper-1-inducing capacity in vitro. We performed the first comparative analysis of the ability of autologous BDCs and MoDCs in healthy donors to induce tumor-specific cytotoxic T lymphocytes (CTLs). BDCs expressed significantly higher levels of major histocompatibility complex class I and CD83 in the absence of exogenous stimuli compared with MoDCs. After activation with polyinosinic-polycytidylic acid, BDCs expressed higher levels of major histocompatibility complex class I, CD40, CD80, and CD83, and secreted higher levels of tumor necrosis factor-alpha, interleukin (IL)-1beta, IL-6, and IL-8 compared with MoDCs. Despite these differences, both preparations secreted similar levels of IL-12 in response to polyinosinic-polycytidylic acid and, importantly, induced CTL responses of similar magnitude and affinity against influenza matrix protein and MART-1. The ability of BDCs to induce efficient CTL responses, combined with their migratory capacity, makes them an appealing alternative to be investigated in clinical immunotherapy research protocols.
Assuntos
Antígeno CD11c/imunologia , Células Dendríticas/imunologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Linfócitos T Citotóxicos/imunologia , Antígenos CD/metabolismo , Antígenos de Diferenciação/metabolismo , Antígenos de Neoplasias/imunologia , Antígeno CD11c/sangue , Ligante de CD40/farmacologia , Linhagem Celular Tumoral , Citocinas/metabolismo , Citocinas/farmacologia , Testes Imunológicos de Citotoxicidade , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/metabolismo , Antígenos HLA-A/imunologia , Antígeno HLA-A2 , Antígenos HLA-DR/metabolismo , Antígenos de Histocompatibilidade Classe I/imunologia , Humanos , Interferon gama/metabolismo , Interleucina-12/metabolismo , Leucócitos Mononucleares/citologia , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/imunologia , Ativação Linfocitária/efeitos dos fármacos , Ativação Linfocitária/imunologia , Antígeno MART-1 , Monócitos/citologia , Monócitos/efeitos dos fármacos , Monócitos/imunologia , Proteínas de Neoplasias/imunologia , Fragmentos de Peptídeos/imunologia , Poli I-C/farmacologia , Linfócitos T Citotóxicos/metabolismo , Receptor 3 Toll-Like/metabolismo , Proteínas da Matriz Viral/imunologiaRESUMO
The lineage of dendritic cells (DC), and in particular their relationship to monocytes and macrophages, remains obscure. Furthermore, the requirement for the macrophage growth factor CSF-1 during DC homeostasis is unclear. Using a transgenic mouse in which the promoter for the CSF-1R (c-fms) directs the expression of enhanced GFP in cells of the myeloid lineage, we determined that although the c-fms promoter is inactive in DC precursors, it is up-regulated in all DC subsets during differentiation. Furthermore, plasmacytoid DC and all CD11c(high) DC subsets are reduced by 50-70% in CSF-1-deficient osteopetrotic mice, confirming that CSF-1 signaling is required for the optimal differentiation of DC in vivo. These data provide additional evidence that the majority of tissue DC is of myeloid origin during steady state and supports a close relationship between DC and macrophage biology in vivo.
Assuntos
Diferenciação Celular/imunologia , Proliferação de Células , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Fator Estimulador de Colônias de Macrófagos/metabolismo , Receptor de Fator Estimulador de Colônias de Macrófagos/biossíntese , Animais , Contagem de Células Sanguíneas , Antígeno CD11c/biossíntese , Antígeno CD11c/sangue , Diferenciação Celular/genética , Células Cultivadas , Células Dendríticas/citologia , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Células-Tronco Hematopoéticas/citologia , Células-Tronco Hematopoéticas/imunologia , Células-Tronco Hematopoéticas/metabolismo , Antígenos Comuns de Leucócito/biossíntese , Fator Estimulador de Colônias de Macrófagos/deficiência , Fator Estimulador de Colônias de Macrófagos/genética , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos CBA , Camundongos Knockout , Camundongos Transgênicos , Receptor de Fator Estimulador de Colônias de Macrófagos/fisiologia , Transdução de Sinais/genética , Transdução de Sinais/imunologiaRESUMO
BACKGROUND: Infants and small children who undergo cardiac surgery due to congenital heart disorders, are at risk of developing inflammatory complications and multi-organ failure. AIM: To assess the changes in the expression of adhesive molecules on neutrophils and in peripheral blood plasma in children undergoing cardiac surgery with or without the use of cardiopulmonary bypass (CPB). METHODS: In 18 children who underwent surgery with the use of CPB and in 7 children who underwent surgery without CPB, the expression of CD11b, CD11c and CD62L molecules on neutrophils and soluble adhesive molecules - L-selectin and ICAM-1, was assessed before, during and after cardiac surgery. RESULTS: A significant increase in the expression of CD11b on neutrophils at the time of the initiation of CPB was observed. The most pronounced decrease in the expression of CD11c was detected at the end of surgical procedure. There was also a negative correlation between L-selectin concentration measured at the end of CPB and CPB duration as well as between ICAM-1 concentration and duration of hypothermia. CONCLUSIONS: Cardiac surgery influences the expression of adhesive molecules on neutrophils and in blood plasma. These changes are more pronounced in children who undergo CPB, and depend on CPB duration as well as the use of hypothermia.
Assuntos
Ponte Cardiopulmonar , Moléculas de Adesão Celular/sangue , Neutrófilos/metabolismo , Antígeno CD11b/sangue , Antígeno CD11c/sangue , Ponte Cardiopulmonar/efeitos adversos , Criança , Pré-Escolar , Feminino , Citometria de Fluxo , Cardiopatias Congênitas/cirurgia , Humanos , Lactente , Molécula 1 de Adesão Intercelular/sangue , Selectina L/sangue , Masculino , Complicações Pós-Operatórias/etiologiaRESUMO
Dendritic cells (DCs) are known to play a major role in the induction, maintenance, and regulation of immune responses. Recently, DCs have been described to be present at the feto-maternal interface in human decidua. However, only limited information is available about DC presence, phenotype, and--more importantly--function throughout gestation. Thus, we analyzed local (uterine) and systemic (blood) DCs in a murine model. DBA/2J mated CBA/J females with vaginal plugs were separated and killed on Gestation Days (GDs) 1.5, 3.5, 5.5, 6.5, 7.5, 8.5, 10.5, 13.5, 15.5, or 17.5. Frequency of uterine and blood CD11c+ DC, phenotype (coexpression of CD8alpha and major histocompatibility complex class II [MHC II] antigens), and presence of intracellular cytokines (interleukins 12 and 10) were determined by flow cytometry. The morphology of DC in the pregnant uterus was evaluated by immunohistochemistry. In uterus, the relative number of CD11c+ cells increased from GD 5.5, reaching a plateau on GD 9.5 until GD 17.5, while a transient peak of systemic CD11c+ cells was found on GD 8.5 and 10.5. The vast majority of uterine DCs were CD8alpha- and thus, belonged to the myeloid lineage. Interestingly, a significant peak of lymphoid DC was present on GD 1.5 and 5.5. Further, significantly more intracellular interleukin 10 than interleukin 12 was present in CD11c+ cells. Interestingly, mature DCs (MHC II+) were diminished from GD 5.5 to 8.5. Characterization of CD11c+ cell kinetics in uterus and blood reveals variation of phenotype during pregnancy, pointing toward an eminent immunoregulatory role of DCs throughout gestation at the feto-maternal interface.
Assuntos
Células Dendríticas/citologia , Células Dendríticas/fisiologia , Prenhez/fisiologia , Útero/citologia , Animais , Células da Medula Óssea/citologia , Antígeno CD11c/sangue , Antígeno CD11c/metabolismo , Linhagem Celular , Senescência Celular , Células Dendríticas/metabolismo , Feminino , Antígenos de Histocompatibilidade Classe II/metabolismo , Tecido Linfoide/citologia , Camundongos , Camundongos Endogâmicos CBA , Camundongos Endogâmicos DBA , Fenótipo , Gravidez , Prenhez/metabolismo , Distribuição Tecidual , Útero/metabolismoRESUMO
The developmental origin of dendritic cells (DCs) is controversial. In the mouse CD8alpha(+) and CD8alpha(-) DC subsets are often considered to be of lymphoid and myeloid origin respectively, although evidence on this point is conflicting. Very recently a novel CD11c(+) B220(+) DC subset has been identified that appears to be the murine counterpart to interferon alpha (IFNalpha)-producing human plasmacytoid DCs (PDCs). We show here that CD11c(+) B220(+) mouse PDCs, like human PDCs, are present in the thymus and express T lineage markers such as CD8alpha and CD4. However, the intrathymic development of PDCs can be completely dissociated from immature T lineage cells in mixed chimeras established with bone marrow cells from mice deficient for either Notch-1 or T-cell factor 1, two independent mutations that severely block early T-cell development. Our data indicate that thymic PDCs do not arise from a bipotential T/DC precursor.
Assuntos
Antígeno CD11c/sangue , Células Dendríticas/imunologia , Antígenos Comuns de Leucócito/sangue , Linfócitos T/imunologia , Animais , Células da Medula Óssea/citologia , Células da Medula Óssea/fisiologia , Quimera/imunologia , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Modelos AnimaisRESUMO
Dendritic cells (DCs) initiate and direct immune responses. Recent studies have defined different DC populations, therefore we undertook this study comparing 2 types of myeloid DCs: blood CD11c(+) DCs and in vitro monocyte-derived DCs (Mo-DCs), which are both candidates as cellular adjuvants for cancer immunotherapy. Blood CD11c(+) DCs were prepared by cell sorting from peripheral blood mononuclear cells cultured overnight in RPMI 1640 medium supplemented with autologous or pooled AB serum. Mo-DCs were prepared in the same medium using granulocyte macrophage-colony-stimulating factor (GM-CSF)/interleukin 4 (IL-4) and differentiated/activated with lipopolysaccharide or monocyte-conditioned medium (ActMo-DCs). Morphologically, differences between the DC preparations were noted both at a light and and electron microscopic level. Blood CD11c(+) DCs expressed similar levels of HLA-DR, CD40, CD86, and CD83 as Mo-DCs. CD209 was present on Mo-DCs but not on blood CD11c(+) DCs. Blood CD11c(+) DCs generated a lower proliferative mixed leukocyte response (MLR) than Mo-DCs. Blood CD11c(+) DCs loaded with 0.1 microg/mL tetanus toxoid (TT)-generated greater T lymphocyte proliferative responses than did Mo-DCs or ActMo-DCs, but when loaded with higher TT concentrations no difference in T lymphocyte proliferative response was observed. Keyhole limpet hemocyanin (KLH)-loaded blood CD11c(+) DCs generated greater T lymphocyte proliferative responses than Mo-DCs or ActMo-DCs. Allogeneic MLR- or KLH-specific responses induced by blood CD11c(+) DCs generated more Th1 effectors than the responses induced by Mo-DCs or ActMo-DCs. These data establish several differences in the properties of blood CD11c(+) DCs, Mo-DCs, and ActMo-DCs, which suggest that blood DCs merit further consideration as DC preparations for clinical programs are evolved.